Proof-of-concept of duplex MNAzyme assay for simultaneous detection of Candida auris and Candida spp. using synthetic DNA targets

Jainlabdin, Mohd Hanif and Ahmed Al-Shaibani, Muhanna Mohammed and Abdel Wahhab, Mosaad Attia and Mohd Ali, Mohammad Ridhuan and Edwards, Thomas and Tovar-Torres, Jorge (2025) Proof-of-concept of duplex MNAzyme assay for simultaneous detection of Candida auris and Candida spp. using synthetic DNA targets. Asia-Pacific Journal of Molecular Biology and Biotechnology, 33 (2). pp. 19-25. ISSN 0128-7451 E-ISSN 2672-7277

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Abstract

The global emergence of Candida auris as a multidrug-resistant fungal pathogen presents significant challenges to healthcare systems, particularly in hospital settings. Rapid and accurate diagnostics are crucial for managing C. auris infections. This study presents a preliminary analytical evaluation of a duplex Multicomponent Nucleic Acid Enzyme (MNAzyme) assay designed to simultaneously detect C. auris and genus-wide Candida species using synthetic DNA targets. Target sequences were selected from the conserved D1/D2 region of the LSU rDNA locus, and melting temperature analysis confirmed distinct probe hybridisation profiles, with Substrate 2:PCO1 yielding a Tm of 45°C and Substrate 3:PCO4 at 26°C. The assay demonstrated specificity by differentiating C. auris from other Candida species through melting curve analysis. In reactions containing synthetic targets, probe– target binding disrupted probe–PCO hybrids, resulting in a marked reduction in melt peaks compared to no DNA controls (NDC). The duplex format allows multiplex detection within a single fluorescence channel, offering a potential advantage over conventional single-target assays. This proof-of-concept establishes a rapid, low-cost diagnostic platform with promising potential for future clinical application. Further validation using clinical and environmental samples is needed to support its broader utility in detecting fungal pathogens.

Item Type:	Article (Journal)
Additional Information:	9313/122588
Uncontrolled Keywords:	C. auris, Candidozyma, One Health, melting curve, multiplex, MNAzymes
Subjects:	Q Science > QR Microbiology R Medicine > RA Public aspects of medicine > RA643 Communicable Diseases and Public Health R Medicine > RT Nursing > RT48 Nursing assessment > RT48.5 Diagnostic techniques, Laboratory tests, etc.
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Nursing > Department of Basic Medical Sciences for Nursing Kulliyyah of Nursing Institute of Planetary Survival for Sustainable Well-Being (PLANETIIUM)
Depositing User:	Dr Mohd Hanif Jainlabdin
Date Deposited:	14 Aug 2025 16:00
Last Modified:	14 Aug 2025 16:01
URI:	http://irep.iium.edu.my/id/eprint/122588

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